Method for Producing Recombinant Stem Bromelain

a technology of stem bromelain and recombinant protein, which is applied in the direction of hydrolases, biochemistry apparatus and processes, enzymes, etc., can solve the problems of high material and operation costs, and achieve the effect of high production rate and cost-effectiveness
US20110151541A1Inactive Publication Date: 2011-06-23INTERNATIONAL ISLAMIC UNIVERSITY MALAYSIA
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Patent Information

Authority / Receiving Office
US · United States
Patent Type
Applications(United States)
Current Assignee / Owner
INTERNATIONAL ISLAMIC UNIVERSITY MALAYSIA
Publication Date
2011-06-23
Estimated Expiration
Not applicable · inactive patent

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Abstract

The present invention relates to a method for producing recombinant stem bromelain. Total RNA is isolated from pineapple plant (11) and used for reverse transcription polymerase chain reaction to generate multiple copies of stem bromelain gene (12). The gene is then cloned into a cloning vector (13), then transformed to a competent cell to produce an entry clone (14). After identifying and selecting the positively transformed entry clone (15), the stem bromelain gene is sub-cloned into a destination vector (16) and transformed into host cells to produce recombinant cells (17). After identifying and selecting the positively transformed recombinant cells (18), the recombinant cells are induced to express recombinant stem bromelain (19). Subsequently, the recombinant cells are harvested and the recombinant stem bromelain purified (20). The recombinant stem bromelain produced is in an active form and demonstrates activity similar to native stem bromelain.
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Description

BACKGROUND OF THE INVENTION

[0001] 1. Field of the Invention

[0002] This invention relates to a method for producing a recombinant protein, and more particularly to a method for producing recombinant stem bromelain.

[0003] 2. Description of Related Arts

[0004] Commercial bromelain is a crude, aqueous extract from the stems and immature fruits of pineapples. Crude pineapple stem preparation contains two distinct cysteine proteases: stem bromelain and ananain. Three cysteine proteases are present in the pineapple stem: stem bromelain, ananain, and comosain. Bromelain is a protein-digesting enzyme similar in function to papain and ficin. It is also the most cost effective natural protease.

[0005] Bromelain is known to have a number of uses. A known pharmaceutical application of bromelain is the enzymic debridement of necrotic tissues from ulcers and burn wounds. Other therapeutic benefits include reversible inhibition of platelet aggregation, management of bronchitis, improved healing of surgica...

Claims

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